Abstract
A direct injection high-performance liquid chromatographic (HPLC) method, with column-switching, for the determination of guaiphenesin (GP) in human serum has been developed. Serum samples were directly injected onto protein-coated RP-8 silica precolumn, where GP was preconcentrated and retained while proteins and very polar components were washed to waste using phosphate buffer saline, pH 7.4. GP was back-flushed from the precolumn by a column-switching technique and separated on a ZORBAX Eclipse XDB-C18 analytical column with a mobile phase consisting of methanol-0.01 M phosphate buffer (containing 1% triethylamine adjusted to pH 3.5 with ortho-phosphoric acid) in the ratio of 45:55 (v/v). The analyte was detected by its UV absorbance at 254 nm. The calibration curve was linear over the concentration range of 25–4000 ng/mL ( r 2 = 0.9994). The method was validated for linearity, accuracy, precision, selectivity, and robustness.
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